RESUMEN
A novel thienopyrimidinone analog was discovered as a potent and highly selective TAK1 inhibitor using the SBDD approach. TAK1 plays a key role in inflammatory and immune signaling, so TAK1 is considered to be an attractive molecular target for the treatment of human diseases (inflammatory disease, cancer, etc.). After the hit compound had been obtained, our modifications successfully increased TAK1 inhibitory activity and solubility, but metabolic stability was still unsatisfactory. To improve metabolic stability, we conducted metabolic identification. Although the obtained metabolite was fortunately a potent TAK1 inhibitor, its kinase selectivity was low. Subsequently, to achieve high kinase selectivity, we used SBDD to follow two strategies: one targeting unique amino acid residues in TAK1, especially the combination of Ser111 and Asn114; the other decreasing the interaction with Tyr106 at the hinge position in TAK1. As expected, our designed compound showed an excellent kinase selectivity profile in both an in-house and a commercially available panel assay of over 420 kinases and also retained its potent TAK1 inhibitory activity (TAK1 IC50=11nM).
Asunto(s)
Quinasas Quinasa Quinasa PAM/antagonistas & inhibidores , Inhibidores de Proteínas Quinasas/farmacología , Pirimidinonas/farmacología , Tiofenos/farmacología , Animales , Asparagina/química , Cristalografía por Rayos X , Diseño de Fármacos , Pruebas de Enzimas , Humanos , Enlace de Hidrógeno , Ratones , Microsomas Hepáticos/efectos de los fármacos , Microsomas Hepáticos/metabolismo , Estructura Molecular , Inhibidores de Proteínas Quinasas/síntesis química , Inhibidores de Proteínas Quinasas/química , Pirimidinonas/síntesis química , Pirimidinonas/química , Serina/química , Solubilidad , Relación Estructura-Actividad , Tiofenos/síntesis química , Tiofenos/química , Tirosina/químicaRESUMEN
Phosphoinositide 3-kinase (PI3K) is a promising anti-cancer target, because various mutations and amplifications are observed in human tumors isolated from cancer patients. Our dihydropyrrolopyrimidine derivative with a phenylurea moiety showed strong PI3K enzyme inhibitory activity, but its pharmacokinetic property was poor because of lack of solubility. Herein, we report how we improved the solubility of our PI3K inhibitors by introducing a solubilizing group and ortho substituents to break molecular planarity.
Asunto(s)
Compuestos de Fenilurea/química , Inhibidores de las Quinasa Fosfoinosítidos-3 , Agua/química , Animales , Activación Enzimática/efectos de los fármacos , Xenoinjertos , Humanos , Concentración 50 Inhibidora , Ratones , Estructura Molecular , Neoplasias/tratamiento farmacológico , Compuestos de Fenilurea/síntesis química , Compuestos de Fenilurea/farmacocinética , Compuestos de Fenilurea/farmacología , SolubilidadRESUMEN
Phosphoinositide 3-kinase (PI3K) is activated in various human cancer cells and well known as a cancer therapy target. We previously reported a dihydropyrrolopyrimidine derivative as a highly potent PI3K inhibitor that has strong tumor growth inhibition in a xenograft model. In this report, we describe further optimization to improve its bioavailability.
Asunto(s)
Inhibidores de las Quinasa Fosfoinosítidos-3 , Inhibidores de Proteínas Quinasas/química , Inhibidores de Proteínas Quinasas/farmacocinética , Pirimidinas/química , Pirimidinas/farmacocinética , Pirroles/química , Pirroles/farmacocinética , Administración Oral , Animales , Disponibilidad Biológica , Femenino , Humanos , Macaca fascicularis , Masculino , Ratones Endogámicos BALB C , Microsomas Hepáticos/metabolismo , Permeabilidad , Inhibidores de Proteínas Quinasas/administración & dosificación , Inhibidores de Proteínas Quinasas/metabolismo , Pirimidinas/administración & dosificación , Pirimidinas/metabolismo , Pirroles/administración & dosificación , Pirroles/metabolismo , SolubilidadRESUMEN
Our lead compound for a phosphoinositide 3-kinase (PI3K) inhibitor (1) was metabolically unstable because of rapid glucuronidation of the phenol moiety. Based on structure-activity relationship (SAR) information and a FlexSIS docking simulation score, aminopyrimidine was identified as a bioisostere of phenol. An X-ray structure study revealed a hydrogen bonding pattern of aminopyrimidine derivatives. Finally, aminopyrimidine derivatives 33 showed strong tumor growth inhibition against a KPL-4 breast cancer xenograft model in vivo.
Asunto(s)
Ácido Glucurónico/química , Fenoles/química , Inhibidores de las Quinasa Fosfoinosítidos-3 , Pirimidinas/síntesis química , Pirroles/síntesis química , Animales , Neoplasias de la Mama/tratamiento farmacológico , Línea Celular Tumoral , Cristalografía por Rayos X , Activación Enzimática/efectos de los fármacos , Femenino , Humanos , Enlace de Hidrógeno , Concentración 50 Inhibidora , Ratones , Estructura Molecular , Pirimidinas/química , Pirimidinas/farmacología , Pirroles/química , Pirroles/farmacología , Relación Estructura-Actividad , Ensayos Antitumor por Modelo de XenoinjertoRESUMEN
Phosphatidylinositol 3-kinase (PI3K) is a lipid kinase and a promising therapeutic target for cancer. Using structure-based drug design (SBDD), we have identified novel PI3K inhibitors with a dihydropyrrolopyrimidine skeleton. Metabolic stability of the first lead series was drastically improved by replacing phenol with aminopyrimidine moiety. CH5132799, a novel class I PI3K inhibitor, exhibited a strong inhibitory activity especially against PI3Kα (IC(50)=0.014 µM). In human tumor cell lines with PI3K pathway activation, CH5132799 showed potent antiproliferative activity. CH5132799 is orally available and showed significant antitumor activity in PI3K pathway-activated human cancer xenograft models in mice.
Asunto(s)
Inhibidores Enzimáticos/farmacología , Inhibidores de las Quinasa Fosfoinosítidos-3 , Pirimidinas/farmacología , Sulfonamidas/farmacología , Línea Celular Tumoral , Humanos , Modelos Moleculares , Fosfatidilinositol 3-Quinasas/químicaRESUMEN
A series of 5,5-dimethylthiohydantoin derivatives were synthesized and evaluated for androgen receptor pure antagonistic activities for the treatment of hormone refractory prostate cancer. CH4933468 (32d) with a sulfonamide side chain not only exhibited antagonistic activity with no agonistic activity in the reporter gene assay but also inhibited the growth of bicalutamide-resistant cell lines. This compound also inhibited tumor growth of the LNCaP xenograft in mice dose-dependently.
Asunto(s)
Antagonistas de Andrógenos , Antineoplásicos Hormonales , Ácidos Carboxílicos , Nitrilos/síntesis química , Sulfonamidas/síntesis química , Antagonistas de Andrógenos/síntesis química , Antagonistas de Andrógenos/química , Antagonistas de Andrógenos/farmacología , Animales , Antineoplásicos Hormonales/síntesis química , Antineoplásicos Hormonales/química , Antineoplásicos Hormonales/farmacología , Ácidos Carboxílicos/síntesis química , Ácidos Carboxílicos/química , Ácidos Carboxílicos/farmacología , Línea Celular Tumoral , Relación Dosis-Respuesta a Droga , Concentración 50 Inhibidora , Masculino , Ratones , Ratones SCID , Estructura Molecular , Nitrilos/química , Nitrilos/uso terapéutico , Neoplasias de la Próstata/tratamiento farmacológico , Sulfonamidas/química , Sulfonamidas/uso terapéutico , Tiohidantoínas/síntesis química , Tiohidantoínas/química , Tiohidantoínas/farmacología , Ensayos Antitumor por Modelo de XenoinjertoRESUMEN
PURPOSE: This case report describes two successful pregnancies after intracytoplasmic sperm injection (ICSI) with testicular spermatozoa that were transported under refrigeration. METHODS: Two first-time couples consulted our clinic concerned about their primary infertility. No sperm were present in the semen samples from either of the husbands and they were referred to the urology department (UD) of a neighbouring hospital. At the UD, seminiferous tubules were obtained by testicular sperm extraction. The tissue samples were put in a centrifuge tube with phosphate-buffered saline at 6°C and placed with refrigerant in a cushioned styrofoam box that was then transported to our clinic. Immediately upon arrival at our clinic, testicular spermatozoa were extracted. On the same day, ovum pickup was performed and mature oocytes were extracted that were then inseminated by conventional ICSI. Fertilized eggs were cultured for 2 days, and then cleaved embryos were cryopreserved. In one case after 4 months and in the other case after 2 months of cryopreservation, the frozen-thawed embryos were transferred. RESULT: Both patients became pregnant and normal, healthy babies were born. CONCLUSIONS: These results suggest that cases of obstructive azoospermia can be treated with ICSI by refrigerated transport of the seminiferous tubules, in cooperation with a UD, in a small single departmental obstetrics and gynecology clinic.
RESUMEN
The 3-(4-cyano-3-trifluoromethylphenyl)-5,5-dimethylthiohydantoin derivatives which have carboxy-terminal side chains were synthesized and their agonistic/antagonistic activities against androgen receptor (AR) measured. Among them, compound 13b showed antagonistic activity (IC50=130 nM) with no agonistic activity even at 10000 nM. This compound exhibited significant metabolic stability and oral antiandrogenic activity (ED50=7 mg/kg).
Asunto(s)
Antagonistas de Andrógenos/síntesis química , Antagonistas de Andrógenos/farmacología , Antagonistas de Receptores Androgénicos , Animales , Unión Competitiva/efectos de los fármacos , Células CHO , Cricetinae , Cricetulus , Cristalografía por Rayos X , Genes Reporteros/efectos de los fármacos , Células HeLa , Humanos , Indicadores y Reactivos , Masculino , Ratones , Ratones Endogámicos ICR , Microsomas Hepáticos/efectos de los fármacos , Microsomas Hepáticos/metabolismo , Modelos Moleculares , Vesículas Seminales/efectos de los fármacos , Relación Estructura-Actividad , Tiohidantoínas/síntesis química , Tiohidantoínas/farmacologíaRESUMEN
Lead optimization of CH4892280 (4), an androgen receptor (AR) pure antagonist, was investigated. Compounds 6 and 7, which have a carboxylic acid at the end of the side chain at the position 7alpha of dihydrotestosterone (DHT), showed partial agonistic activities in reporter gene assay (RGA). Conversion of the steroidal core structure to 17alpha-methyltestosterone gave compound 14, which showed weak pure antagonistic activity. Optimization of the side chain by the insertion of a phenyl ring led to compounds 22 and 28-30, which showed pure antagonistic activities at submicromolar concentrations. The structure-activity relationships were clarified.
Asunto(s)
Antagonistas de Receptores Androgénicos , Receptores Androgénicos/metabolismo , Esteroides/química , Esteroides/farmacología , Animales , Ratones , Modelos Moleculares , Estructura Molecular , Unión Proteica , Receptores Androgénicos/química , Esteroides/síntesis química , Relación Estructura-ActividadRESUMEN
A series of 7alpha-substituted dihydrotestosterone derivatives were synthesized and evaluated for androgen receptor (AR) pure antagonistic activity. From reporter gene assay (RGA), the compound with a side chain containing N-n-butyl-N-methyl amide (19a) showed pure antagonistic activity (IC(50)=340nM, FI(5)>10,000nM), whereas known AR antagonists showed partial agonistic activities. The optimization of 19a led to compound 23 (CH4892280), which showed more potent pure antagonistic activity (IC(50)=190nM, FI(5)>10,000nM). The SARs of tested compounds suggested that the length of the side chain and the substituents on the amide nitrogen are important for pure antagonistic activities.
Asunto(s)
Antagonistas de Receptores Androgénicos , Dihidrotestosterona/análogos & derivados , Dihidrotestosterona/farmacología , Animales , Sitios de Unión , Unión Competitiva/efectos de los fármacos , Células CHO , Cricetinae , Dihidrotestosterona/química , Relación Dosis-Respuesta a Droga , Evaluación Preclínica de Medicamentos , Células HeLa , Humanos , Estructura Molecular , Estereoisomerismo , Relación Estructura-ActividadRESUMEN
A small, time efficient and sensitive instrument for the continuous analysis of very volatile organic compounds (VOCs) with a boiling point lower than 100 degrees C in addition to the analysis of VOCs with a boiling point in the range of 100-150 degrees C was developed and applied to the measurement of VOCs in the course of university research and environmental monitoring. VOCs, such as n-hexane, acetone, ethyl acetate, alcohols, benzene, toluene and xylene, were continuously measured once every 30 min. The detection limits of hexane, ethyl acetate, benzene and toluene at a preconcentration time of 10 min were 0.41 microg/m(3) (0.12 ppb), 0.67 microg/m(3) (0.19 ppb), 0.22 microg/m(3) (0.07 ppb) and 0.22 microg/m(3) (0.06 ppb), respectively. The relative standard deviations of VOCs were less than 5%. The sensitivities of the present method VOCs were higher than those of the conventional method. The temporal changes in VOC concentrations in several laboratories and at a plant for the disposal of organic liquid wastes were measured, and the behavior of VOCs was analyzed. All the VOC concentrations, except that of ethyl acetate, determined using the portable instrument were slightly lower than those determined using a passive sampler. The portable instrument developed in the course of this study can be used for the risk assessment and management of chemicals.
Asunto(s)
Contaminantes Atmosféricos/análisis , Monitoreo del Ambiente/instrumentación , Compuestos Orgánicos/análisis , Diseño de Equipo , Temperatura de Transición , VolatilizaciónRESUMEN
Tissue factor (TF) is a membrane-anchored protein that initiates the extrinsic cascade of blood coagulation. TF forms a complex with serine protease Factor VIIa, and then activates Factor X zymogen to Factor Xa, leading to the blood coagulation. Humanized anti-TF antibody hATR-5 strongly inhibits TF-initiated blood coagulation, and is of potential use for various thrombotic diseases. The Fab fragment of antibody hATR-5 is obtained for crystallization. The crystal structure of the complex of the Fab with extracellular domains of human TF was determined with the molecular replacement method, and refined to an R factor of 0.196 at 2.1 A resolution. All the complementarity-determining regions (CDRs) of the Fab are involved in interaction with the C-terminal-side extracellular domain of TF through 19 hydrogen bonds. The interface between the Fab and TF molecules contains 15 water molecules, and yields buried surface areas as wide as 2000 A2. The TF surface in the interface is possibly involved in the activation of Factor X, by forming a transient ternary complex of Factor X-TF-Factor VIIa. Electrostatic interactions are predominantly observed between the heavy-chain CDRs and TF. These hydrogen-bonding and electrostatic interactions together with the wide buried areas contribute to the high affinity of the antibody toward TF, leading to the effective inhibition of the TF-initiated blood coagulation.